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Evaluation of FKBP and DHFR based destabilizing domains in Saccharomyces cerevisiae.


ABSTRACT: Two orthogonal destabilizing domains have been developed based on mutants of human FKBP12 as well as bacterial DHFR and these engineered domains have been used to control protein concentration in a variety of contexts in vitro and in vivo. FKBP12 based destabilizing domains cannot be rescued in the yeast Saccharomyces cerevisiae; ecDHFR based destabilizing domains are not degraded as efficiently in S. cerevisiae as in mammalian cells or Plasmodium, but provide a starting point for the development of domains with increased signal-to-noise in S. cerevisiae.

SUBMITTER: Rakhit R 

PROVIDER: S-EPMC3156383 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature

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Evaluation of FKBP and DHFR based destabilizing domains in Saccharomyces cerevisiae.

Rakhit Rishi R   Edwards Sarah R SR   Iwamoto Mari M   Wandless Thomas J TJ  

Bioorganic & medicinal chemistry letters 20110616 17


Two orthogonal destabilizing domains have been developed based on mutants of human FKBP12 as well as bacterial DHFR and these engineered domains have been used to control protein concentration in a variety of contexts in vitro and in vivo. FKBP12 based destabilizing domains cannot be rescued in the yeast Saccharomyces cerevisiae; ecDHFR based destabilizing domains are not degraded as efficiently in S. cerevisiae as in mammalian cells or Plasmodium, but provide a starting point for the developmen  ...[more]

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